Cencic Regina, Senechal Patrick, Pelletier Jerry
Rm 810, 3655 Promenade Sir William Osler., McIntyre Medical Sciences Building, Montreal, Quebec, H3G 1Y6, Canada.
Comb Chem High Throughput Screen. 2015;18(9):855-61. doi: 10.2174/1386207318666151019093433.
The RNA helicase DHX9 is an ATP-dependent DExH box helicase that can unwind DNA and RNA. Much evidence has implicated DHX9 at multiple levels of gene expression regulation ranging from genome stability and replication, to transcriptional control and translation regulation. Its association with the EWS-FLI1 fusion product, as well as the finding that its suppression can be synthetic lethal with the BCL-2 family inhibitor ABT-737 indicates a potential role in tumor maintenance. Hence, to identify small molecules that could interfere with its activity, we developed a homogenous RNA-dependent ATPase assay. We show that aurintricarboxylic acid, a promiscuous protein-nucleic acid inhibitor prevents DHX9-mediated hydrolysis demonstrating that the assay is also capable of detecting compounds that impinge on DHX9:RNA association.
RNA解旋酶DHX9是一种依赖ATP的DExH盒解旋酶，能够解开DNA和RNA。大量证据表明，DHX9在基因表达调控的多个层面发挥作用，范围从基因组稳定性和复制，到转录控制和翻译调控。它与EWS-FLI1融合产物相关联，并且其抑制作用与BCL-2家族抑制剂ABT-737具有合成致死性这一发现表明它在肿瘤维持中具有潜在作用。因此，为了鉴定能够干扰其活性的小分子，我们开发了一种均相RNA依赖性ATP酶检测方法。我们发现，金精三羧酸这种混杂的蛋白质-核酸抑制剂可阻止DHX9介导的水解反应，这表明该检测方法也能够检测影响DHX9与RNA结合的化合物。
